Fig. 4.

Dual color FACS for identification of inhibitory antibodies and epitope profiling. (A) Antibody-displaying cells are incubated with Alexa488-cdMMP14 and Alexa647-nTIMP2 sequentially, and subjected to FACS scanning. Three scenarios are expected: non-specific, specific but non-inhibitory and inhibitory antibodies. (B) Distinguishment of binding and inhibitory clones by dual color FACS. Non-specific antibody M18 (anti-PA) and specific but non-inhibitory antibody 2A10 are used as negative controls. DX-2400, inhibitory Ab directly competing with n-TIMP-2, is used as a positive control. 1F8 competes with n-TIMP-2 on MMP-14 inhibition.